Selected technologies to control genes and their products for experimental and clinical purposes

  • Helen K. Alexander
  • Evan P. Booy
  • Wenyan Xiao
  • Peyman Ezzati
  • Heinrich Baust
  • Marek Los
Review

Abstract.

“On-demand” regulation of gene expression is a powerful tool to elucidate the functions of proteins and biologically-active RNAs. We describe here three different approaches to the regulation of expression or activity of genes or proteins. Promoter-based regulation of gene expression was among the most rapidly developing techniques in the 1980s and 1990s. Here we provide basic information and also some characteristics of the metallothionein-promoter-based system, the tet-off system, Muristerone-A-regulated expression through the ecdysone response element, RheoSwitch®, coumermycin/novobiocin-regulated gene expression, chemical dimerizer-based promoter activation systems, the “Dual Drug Control” system, “constitutive androstane receptor” based regulation of gene expression, and RU486/mifepristone-driven regulation of promoter activity. A large part of the review concentrates on the principles and usage of various RNA interference techniques (RNAi: siRNA, shRNA, and miRNA-based methods). Finally, the last part of the review deals with historically the oldest, but still widely used, methods of temperature-dependent regulation of enzymatic activity or protein stability (temperature-sensitive mutants). Due to space limitations we do not describe in detail but just mention the tet-regulated systems and also fusion-protein-based regulation of protein activity, such as estrogen-receptor fusion proteins. The information provided below is aimed to assist researchers in choosing the most appropriate method for the planned development of experimental systems with regulated expression or activity of studied proteins.

Keywords:

Dicer Drosha FKBP gene silencing Mifepristone mutagensis N-degron NF-κB RISC 

Abbreviations:

CAR

constitutive androstane receptor

dsRNA

double-stranded RNA

DHFR

dihydrofolate reductase

EcR

ecdysone receptor

FKBP

FK506-binding protein

FRAP

FKBP-rapamycin-associated protein

Hsp-90

heat shock protein 90

MRE

metal-responsive elements

miRNAs

microRNAs

MCM

minichromosome maintenance

NF-κB

nuclear factor κB

PBREM

phenobarbital-responsive enhancer module

tetR

tetracycline-dependent repressor

tTA

tetracycline-controlled transactivator

tetO

tetracycline operator

TK

thymidine kinase

RXR

retinoid X receptor

RNAi

RNA interference

RISC

RNA-induced silencing complex

shRNAs

short hairpin RNAs

siRNAs

small interfering RNAs

td

temperature-inducible degron

ts

temperature-sensitive

VP16

virion protein 16 of herpes simplex virus

Copyright information

© Birkhäuser Verlag, Basel 2007

Authors and Affiliations

  • Helen K. Alexander
    • 1
  • Evan P. Booy
    • 1
    • 3
  • Wenyan Xiao
    • 1
  • Peyman Ezzati
    • 1
  • Heinrich Baust
    • 4
  • Marek Los
    • 1
    • 2
    • 3
  1. 1.Cancer Care ManitobaManitoba Institute of Cell Biology, University of ManitobaWinnipegCanada
  2. 2.Department of Human Anatomy and Cell ScienceUniversity of ManitobaWinnipegCanada
  3. 3.Department of Biochemistry and Medical GeneticsUniversity of ManitobaWinnipegCanada
  4. 4.Department of RadiooncologyUniversity of ErlangenErlangenGermany

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